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Scientific article
English

Regulation of tumor necrosis factor alpha transcription in macrophages: involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B

Published inMolecular and cellular biology, vol. 10, no. 4, p. 1498-1506
Publication date1990
Abstract

This study characterizes the interaction of murine macrophage nuclear proteins with the tumor necrosis factor alpha (TNF-alpha) promoter. Gel retardation and methylation interference assays showed that stimulation of TNF-alpha gene transcription in peritoneal exudate macrophages was accompanied by induction of DNA-binding proteins that recognized with different affinities four elements related to the kappa B consensus motif and a Y-box motif. We suggest that the basal level of TNF-alpha expression in macrophages is due to the binding of a constitutive form of NF-kappa B, present at low levels in nuclei from resting thioglycolate exudate peritoneal macrophages, to some if not all of the kappa B motifs; we postulate that this constitutive form contains only the 50-kilodalton (kDa) DNA-binding protein subunits of NF-kappa B, not the 65-kDa protein subunits (P. Baeuerle and D. Baltimore, Genes Dev. 3:1689-1698, 1989). Agents such as glucocorticoids, which decrease TNF-alpha transcription, diminished the basal level of nuclear NF-kappa B. Stimulation of Stimulation of TNF-alpha transcription in macrophages by lipopolysaccharide, gamma interferon, or cycloheximide led to an increased content of nuclear NF-kappa B. This induced factor represents a different form of NF-kappa B, since it generated protein-DNA complexes of slower mobility; we propose that this induced form of NF-kappa B contains both the 50- and 65-kDa protein subunits, the latter ones being necessary to bind NF-kappa B to its cytoplasmic inhibitor in uninduced cells (Baeuerle and Baltimore, Genes Dev., 1989). In resting cells, this inducible form of NF-kappa B was indeed detectable in the cytosol after deoxycholate treatment. UV cross-linking experiments and gel retardation assays indicated that the inducible form of NF-kappa B is in a higher-order complex with other proteins.

Keywords
  • Animals
  • Base Sequence
  • Cell Nucleus/metabolism
  • Cells, Cultured
  • Cycloheximide/pharmacology
  • DNA Probes
  • Gene Expression Regulation
  • Lipopolysaccharides/pharmacology
  • Macrophage Activation
  • Macrophages/drug effects/physiology
  • Mice
  • Mice, Inbred CBA
  • Molecular Sequence Data
  • NF-kappa B
  • Nuclear Proteins/biosynthesis/metabolism
  • Promoter Regions, Genetic
  • Templates, Genetic
  • Transcription Factors/genetics/metabolism
  • Transcription, Genetic
  • Tumor Necrosis Factor-alpha/genetics
Citation (ISO format)
COLLART, Martine, BAEUERLE, P, VASSALLI, Pierre. Regulation of tumor necrosis factor alpha transcription in macrophages: involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B. In: Molecular and cellular biology, 1990, vol. 10, n° 4, p. 1498–1506. doi: 10.1128/mcb.10.4.1498
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Article (Published version)
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Identifiers
ISSN of the journal0270-7306
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